Unit 1.1.c Flashcards

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1
Q

What is the use of a centrifuge?

A

To separate substances of differing density

More dense components settle in the pellet; less dense components remain in the supernatant.

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2
Q

What can paper and thin chromatography be used for

A

For separating different substances such as amino acids and sugars

The speed that each solute travels along the chromatogram depends on its differing solubility in the solvent used.

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3
Q

What is affinity chromatography used for?

A

Separating proteins

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4
Q

What is the principle of affinity chromatography

A

A solid matrix or gel column is created with specific molecules bound to the matrix or gel. Soluble, target proteins in a mixture, with a high affinity for these molecules, become attached to them as the mixture passes down the column. Other non-target molecules with a weaker affinity are washed out

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5
Q

What is gel electrophoresis used for?

A

Separating proteins and nucleic acids

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6
Q

What is the principle of gel electrophoresis?

A

Charged macromolecules move through an electric field applied to a gel matrix.

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7
Q

How do native gels separate proteins?

A

Native gels do not denature the molecule so that separation is by shape, size and charge

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8
Q

How does SDS-PAGE separate proteins?

A

SDS–PAGE gives all the molecules an equally negative charge and denatures them, separating proteins by size alone.

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9
Q

What is IEP and what is it used for?

A

IEP is the pH at which a soluble protein has no net charge and will precipitate out of solution.

Proteins can be separated from a mixture using their isoelectric points (IEPs)

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10
Q

What happens if the solution is buffered to a specific pH?

A

only the protein(s) that have an IEP of that pH will precipitate

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11
Q

How can proteins be separated using their IEPs in electrophoresis?

A

Soluble proteins can be separated using an electric field and a pH gradient. A protein stops migrating through the gel at its IEP in the pH gradient because it has no net charge.

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